CN101424682B - Biological chip base processing method - Google Patents
Biological chip base processing method Download PDFInfo
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- CN101424682B CN101424682B CN 200710134477 CN200710134477A CN101424682B CN 101424682 B CN101424682 B CN 101424682B CN 200710134477 CN200710134477 CN 200710134477 CN 200710134477 A CN200710134477 A CN 200710134477A CN 101424682 B CN101424682 B CN 101424682B
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- purified water
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- 238000003672 processing method Methods 0.000 title claims description 5
- 239000008213 purified water Substances 0.000 claims abstract description 26
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 26
- 238000006243 chemical reaction Methods 0.000 claims abstract description 21
- 238000001035 drying Methods 0.000 claims abstract description 18
- 239000012530 fluid Substances 0.000 claims abstract description 18
- 229910021642 ultra pure water Inorganic materials 0.000 claims abstract description 18
- 239000012498 ultrapure water Substances 0.000 claims abstract description 18
- 238000007037 hydroformylation reaction Methods 0.000 claims abstract description 17
- 238000005406 washing Methods 0.000 claims abstract description 10
- 239000000243 solution Substances 0.000 claims description 56
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 claims description 24
- RFSUNEUAIZKAJO-ARQDHWQXSA-N Fructose Chemical compound OC[C@H]1O[C@](O)(CO)[C@@H](O)[C@@H]1O RFSUNEUAIZKAJO-ARQDHWQXSA-N 0.000 claims description 17
- 229930091371 Fructose Natural products 0.000 claims description 17
- 239000005715 Fructose Substances 0.000 claims description 17
- 206010013786 Dry skin Diseases 0.000 claims description 14
- SXRSQZLOMIGNAQ-UHFFFAOYSA-N Glutaraldehyde Chemical compound O=CCCCC=O SXRSQZLOMIGNAQ-UHFFFAOYSA-N 0.000 claims description 9
- WYTZZXDRDKSJID-UHFFFAOYSA-N (3-aminopropyl)triethoxysilane Chemical compound CCO[Si](OCC)(OCC)CCCN WYTZZXDRDKSJID-UHFFFAOYSA-N 0.000 claims description 8
- 239000007864 aqueous solution Substances 0.000 claims description 8
- 238000011010 flushing procedure Methods 0.000 claims description 8
- 238000003756 stirring Methods 0.000 claims description 2
- 238000002360 preparation method Methods 0.000 claims 1
- 230000000694 effects Effects 0.000 abstract description 9
- 239000012472 biological sample Substances 0.000 abstract description 7
- 239000000523 sample Substances 0.000 abstract description 7
- 238000000034 method Methods 0.000 abstract description 4
- 238000000018 DNA microarray Methods 0.000 abstract description 3
- 239000012620 biological material Substances 0.000 abstract description 2
- 238000001816 cooling Methods 0.000 abstract 3
- 230000004913 activation Effects 0.000 abstract 2
- 238000009432 framing Methods 0.000 abstract 2
- 238000007654 immersion Methods 0.000 abstract 2
- 238000007605 air drying Methods 0.000 abstract 1
- -1 air drying Chemical compound 0.000 abstract 1
- 239000000758 substrate Substances 0.000 description 7
- 238000004519 manufacturing process Methods 0.000 description 5
- 102000013529 alpha-Fetoproteins Human genes 0.000 description 3
- 108010026331 alpha-Fetoproteins Proteins 0.000 description 3
- 230000003247 decreasing effect Effects 0.000 description 3
- 239000011521 glass Substances 0.000 description 3
- 102000008102 Ankyrins Human genes 0.000 description 2
- 108010049777 Ankyrins Proteins 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- QKNYBSVHEMOAJP-UHFFFAOYSA-N 2-amino-2-(hydroxymethyl)propane-1,3-diol;hydron;chloride Chemical compound Cl.OCC(N)(CO)CO QKNYBSVHEMOAJP-UHFFFAOYSA-N 0.000 description 1
- 238000002965 ELISA Methods 0.000 description 1
- 108010001336 Horseradish Peroxidase Proteins 0.000 description 1
- BLRPTPMANUNPDV-UHFFFAOYSA-N Silane Chemical compound [SiH4] BLRPTPMANUNPDV-UHFFFAOYSA-N 0.000 description 1
- XUIMIQQOPSSXEZ-UHFFFAOYSA-N Silicon Chemical compound [Si] XUIMIQQOPSSXEZ-UHFFFAOYSA-N 0.000 description 1
- 239000000427 antigen Substances 0.000 description 1
- 102000036639 antigens Human genes 0.000 description 1
- 108091007433 antigens Proteins 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 244000309466 calf Species 0.000 description 1
- 239000006210 lotion Substances 0.000 description 1
- 238000002203 pretreatment Methods 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 230000035945 sensitivity Effects 0.000 description 1
- 210000002966 serum Anatomy 0.000 description 1
- 229910000077 silane Inorganic materials 0.000 description 1
Landscapes
- Immobilizing And Processing Of Enzymes And Microorganisms (AREA)
Abstract
The invention belongs to the field of biomaterial and provides a method for processing a biochip base, which can reduce the framing effect of the chip base and can provide high degree of precision meeting the requirement of clinical use, high binding capacity of a biological sample and the chip base and slow activation depress of a fixed sample. The method adopts the technical proposal of preparing a treatment fluid for the chip base and immersing and processing the chip base in the treatment fluid at room temperature and then processes the chip base by the steps of ultrasonic washing with purified water, air drying, KOH immersion, room temperature reaction, purified water rinsing, drying at 60 DEG C, cooling to the room temperature, silicification solution immersion, lucifugal reaction at the room temperature, ultrapure water rinsing, drying and cooling to the room temperature. Then the silicification chip base is immersed in and reacts with a hydroformylation solution at a temperature from 30 to 45 DEG C and is processed by the steps of rinsing by the ultrapure water for six times, 10-40 minute drying at 110 DEG C and cooling to the room temperature. The produced biochip base has the advantages of low framing effect, high degree of precision, slow activation depress of the fixed sample and high binding capacity of the biological sample and the chip base.
Description
Technical field
The invention belongs to technical field of biological material.
Background technology
At present, the making of the biochip of hard substrate form, mainly be with hard substrates such as glass sheet or silicon chips after chemical conversion treatment, direct synthesising probing needle on substrate, perhaps with synthetic probe, antibody, the biological samples such as antigen are selected and are added on the substrate, react by chemical bond, biological sample is fixed on the hard substrate, its advantage is the sample fixation, precision is good, but its shortcoming is that glass sheet sheet base frame effect is obvious, and precision still fails to satisfy clinical request for utilization, biological sample and sheet base binding capacity are low, and the fixed sample activity decreased is fast etc.
Summary of the invention
Technical matters to be solved by this invention is: provide a kind of and can reduce chip slapper base frame effect, precision can satisfy clinical request for utilization, and biological sample and sheet base binding capacity are high, the biological chip base processing method that the fixed sample activity decreased is slow.
Technical scheme of the present invention is:
(1) with H
2SO
4With H
2O
250%-80% by volume: 20%-50% stirs and evenly mixs, and is made into chip slapper base treating fluid;
(2) the chip slapper base is immersed in the sheet base treating fluid room temperature treatment 12-36 hour;
(3) take out purified water supersound washing 5 times, 10min/ time;
(4) air-dry;
(5) immerse 5%-30%KOH solution, room temperature reaction 30-120 minute;
(6) take out with purified water flushing 3 times;
(7) 60 degrees centigrade of oven dry;
(8) be cooled to room temperature;
(9) the 3-aminopropyl triethoxysilane is made into 1%-12% solution with purified water after, add 0.1% fructose and be made into silication solution;
(10) immerse in the silication solution room temperature lucifuge reaction 30 minutes;
(11) take out with ultrapure water rinsing 2 times;
(12) 120-180 degree centigrade of drying is 30 minutes;
(13) be cooled to room temperature;
(14) glutaraldehyde is made into adds 1% fructose after the 1%-10% aqueous solution, transfer pH to 8.0-10 with 5%NaOH solution, make hydroformylation solution;
(15) silication sheet base is entered hydroformylation solution, 30-45 degree centigrade was reacted 10-90 minute;
(16) the ultrapure water rinsing is 6 times;
(17) 110 degrees centigrade dry 10-40 minute;
(18) be cooled to room temperature.
Beneficial effect: biological chip base processing method of the present invention, by in the pre-treatment of sheet base, adding H
2SO
4With H
2O
2And the KOH effects on surface carries out pre-service, adding fructose and suitable silane concentration, glutaraldehyde concentration and appropriate high-temperature baking in the surface active process, the biological chip base of making, the frame effect is low, precision is high, and the fixed sample activity decreased is slow, and biological sample and sheet base binding capacity are high.
Embodiment
The present invention is described in detail below in conjunction with embodiment.
Embodiment 1
(1) with H
2SO
4With H
2O
2Stirred and evenly mixed in 50%: 50% by volume, and be made into sheet base treating fluid
(2) immerse in the sheet base treating fluid room temperature treatment 12 hours
(3) take out purified water supersound washing 5 times, 10min/ time
(4) air-dry
(5) immerse 5%KOH solution, room temperature reaction 30 minutes
(6) take out with purified water flushing 3 times
(7) 60 degrees centigrade of oven dry
(8) be cooled to room temperature
(9) the 3-aminopropyl triethoxysilane is made into 1% solution with purified water after, add 0.1% fructose and be made into silication solution
(10) immerse in the silication solution room temperature lucifuge reaction 30 minutes
(11) take out with ultrapure water rinsing 2 times
(12) 120 degrees centigrade of dryings 30 minutes
(13) be cooled to room temperature
(14) glutaraldehyde is made into adds 1% fructose after 1% aqueous solution, transfer pH to 8.0 with 5%NaOH solution, make hydroformylation solution
(15) silication sheet base is entered hydroformylation solution, 30 degrees centigrade were reacted 10 minutes
(16) the ultrapure water rinsing is 6 times
(17) 110 degrees centigrade of dryings 10 minutes
(18) be cooled to room temperature
Embodiment 2
(1) with H
2SO
4With H
2O
2Stirred and evenly mixed in 55%: 45% by volume, and be made into sheet base treating fluid
(2) immerse in the sheet base treating fluid room temperature treatment 16 hours
(3) take out purified water supersound washing 5 times, 10min/ time
(4) air-dry
(5) immerse 10%KOH solution, room temperature reaction 50 minutes
(6) take out with purified water flushing 3 times
(7) 60 degrees centigrade of oven dry
(8) be cooled to room temperature
(9) the 3-aminopropyl triethoxysilane is made into 3% solution with purified water after, add 0.1% fructose and be made into silication solution
(10) immerse in the silication solution room temperature lucifuge reaction 30 minutes
(11) take out with ultrapure water rinsing 2 times
(12) 130 degrees centigrade of dryings 30 minutes
(13) be cooled to room temperature
(14) glutaraldehyde is made into adds 1% fructose after 2% aqueous solution, transfer pH to 8.2 with 5%NaOH solution, make hydroformylation solution
(15) silication sheet base is entered hydroformylation solution, 34 degrees centigrade were reacted 20 minutes
(16) the ultrapure water rinsing is 6 times
(17) 110 degrees centigrade of dryings 15 minutes
(18) be cooled to room temperature
Embodiment 3
(1) with H
2SO
4With H
2O
2Stirred and evenly mixed in 60%: 40% by volume, and be made into sheet base treating fluid
(2) immerse in the sheet base treating fluid room temperature treatment 20 hours
(3) take out purified water supersound washing 5 times, 10min/ time
(4) air-dry
(5) immerse 15%KOH solution, room temperature reaction 70 minutes
(6) take out with purified water flushing 3 times
(7) 60 degrees centigrade of oven dry
(8) be cooled to room temperature
(9) the 3-aminopropyl triethoxysilane is made into 5% solution with purified water after, add 0.1% fructose and be made into silication solution
(10) immerse in the silication solution room temperature lucifuge reaction 30 minutes
(11) take out with ultrapure water rinsing 2 times
(12) 140 degrees centigrade of dryings 30 minutes
(13) be cooled to room temperature
(14) glutaraldehyde is made into adds 1% fructose after 3% aqueous solution, transfer pH to 8.5 with 5%NaOH solution, make hydroformylation solution
(15) silication sheet base is entered hydroformylation solution, 38 degrees centigrade were reacted 30 minutes
(16) the ultrapure water rinsing is 6 times
(17) 110 degrees centigrade of dryings 20 minutes
(18) be cooled to room temperature
Embodiment 4
(1) with H
2SO
4With H
2O
2Stirred and evenly mixed in 65%: 35% by volume, and be made into sheet base treating fluid
(2) immerse in the sheet base treating fluid room temperature treatment 24 hours
(3) take out purified water supersound washing 5 times, 10min/ time
(4) air-dry
(5) immerse 20%KOH solution, room temperature reaction 90 minutes
(6) take out with purified water flushing 3 times
(7) 60 degrees centigrade of oven dry
(8) be cooled to room temperature
(9) the 3-aminopropyl triethoxysilane is made into 7% solution with purified water after, add 0.1% fructose and be made into silication solution
(10) immerse in the silication solution room temperature lucifuge reaction 30 minutes
(11) take out with ultrapure water rinsing 2 times
(12) 150 degrees centigrade of dryings 30 minutes
(13) be cooled to room temperature
(14) glutaraldehyde is made into adds 1% fructose after 5% aqueous solution, transfer pH to 9.0 with 5%NaOH solution, make hydroformylation solution
(15) silication sheet base is entered hydroformylation solution, 40 degrees centigrade were reacted 50 minutes
(16) the ultrapure water rinsing is 6 times
(17) 110 degrees centigrade of dryings 30 minutes
(18) be cooled to room temperature
Embodiment 5
(1) with H
2SO
4With H
2O
2Stirred and evenly mixed in 70%: 30% by volume, and be made into sheet base treating fluid
(2) immerse in the sheet base treating fluid room temperature treatment 28 hours
(3) take out purified water supersound washing 5 times, 10min/ time
(4) air-dry
(5) immerse 25%KOH solution, room temperature reaction 110 minutes
(6) take out with purified water flushing 3 times
(7) 60 degrees centigrade of oven dry
(8) be cooled to room temperature
(9) the 3-aminopropyl triethoxysilane is made into 9% solution with purified water after, add 0.1% fructose and be made into silication solution
(10) immerse in the silication solution room temperature lucifuge reaction 30 minutes
(11) take out with ultrapure water rinsing 2 times
(12) 160 degrees centigrade of dryings 30 minutes
(13) be cooled to room temperature
(14) glutaraldehyde is made into adds 1% fructose after 7% aqueous solution, transfer pH to 9.5 with 5%NaOH solution, make hydroformylation solution
(15) silication sheet base is entered hydroformylation solution, 42 degrees centigrade were reacted 70 minutes
(16) the ultrapure water rinsing is 6 times
(17) 110 degrees centigrade of dryings 35 minutes
(18) be cooled to room temperature
Embodiment 6
(1) with H
2SO
4With H
2O
2Stirred and evenly mixed in 80%: 20% by volume, and be made into sheet base treating fluid
(2) immerse in the sheet base treating fluid room temperature treatment 36 hours
(3) take out purified water supersound washing 5 times, 10min/ time
(4) air-dry
(5) immerse 30%KOH solution, room temperature reaction 120 minutes
(6) take out with purified water flushing 3 times
(7) 60 degrees centigrade of oven dry
(8) be cooled to room temperature
(9) the 3-aminopropyl triethoxysilane is made into 12% solution with purified water after, add 0.1% fructose and be made into silication solution
(10) immerse in the silication solution room temperature lucifuge reaction 30 minutes
(11) take out with ultrapure water rinsing 2 times
(12) 180 degrees centigrade of dryings 30 minutes
(13) be cooled to room temperature
(14) glutaraldehyde is made into adds 1% fructose after 10% aqueous solution, transfer pH to 10.0 with 5%NaOH solution, make hydroformylation solution
(15) silication sheet base is entered hydroformylation solution, 45 degrees centigrade were reacted 90 minutes
(16) the ultrapure water rinsing is 6 times
(17) 110 degrees centigrade of dryings 40 minutes
(18) be cooled to room temperature
Table 1 is the part of detecting result of embodiment of the invention 1-6
The raw material that is used for test in the table 1 is respectively:
Ankyrin: AFP (Alpha-Fetoprotein) antibody (the rich match in Henan biological production)
Signal antibody: horseradish peroxidase-labeled AFP antibody (the rich match in Henan biological production)
Sealer: calf serum (the general imperial biological production in Shanghai)
Reaction washing lotion: 0.1mol/l Tris-Hcl (biological production of U.S. season of Shanghai)
Luminous substrate: SuperSignal ELISA Femto Maximum Sensitivity Substrate (U.S. pierce production)
Specking spotting robot with the U.S. biodot company during test is put ankyrin after surface of glass slide by the 4*12 array way, through the reaction read signal, analyzes.
Table 1: test result
Claims (1)
1. a biological chip base processing method is characterized in that, comprises the steps:
(1) with H
2SO
4With H
2O
250%-80% by volume: 20%-50% stirs and evenly mixs, and is made into sheet base treating fluid;
(2) the chip slapper base is immersed in the sheet base treating fluid room temperature treatment 12-36 hour;
(3) take out purified water supersound washing 5 times, 10min/ time;
(4) air-dry;
(5) immerse 5%-30%KOH solution, room temperature reaction 30-120 minute;
(6) take out with purified water flushing 3 times;
(7) 60 degrees centigrade of oven dry;
(8) be cooled to room temperature;
(9) the 3-aminopropyl triethoxysilane is made into 1%-12% solution with purified water after, add 0.1% fructose and be made into silication solution;
(10) will immerse through the biological chip base that step 8 is processed by in the step 9 preparation silication solution room temperature lucifuge reaction 30 minutes;
(11) take out with ultrapure water rinsing 2 times;
(12) 120 degrees centigrade of dryings 30 minutes;
(13) be cooled to room temperature;
(14) glutaraldehyde is made into adds 1% fructose after the 1%-10% aqueous solution, transfer pH to 8.0-10 with 5%NaOH solution, make hydroformylation solution;
(15) silication sheet base is immersed hydroformylation solution, 30-45 degree centigrade was reacted 10-90 minute;
(16) the ultrapure water rinsing is 6 times;
(17) 110 degrees centigrade of dryings were divided 10-40 minute;
(18) be cooled to room temperature.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 200710134477 CN101424682B (en) | 2007-10-30 | 2007-10-30 | Biological chip base processing method |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 200710134477 CN101424682B (en) | 2007-10-30 | 2007-10-30 | Biological chip base processing method |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN101424682A CN101424682A (en) | 2009-05-06 |
| CN101424682B true CN101424682B (en) | 2013-01-02 |
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| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN 200710134477 Active CN101424682B (en) | 2007-10-30 | 2007-10-30 | Biological chip base processing method |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN101424682B (en) |
Families Citing this family (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108387750A (en) * | 2018-02-08 | 2018-08-10 | 江苏三联生物工程有限公司 | A kind of chip and preparation method thereof of joint-detection F-T3, F-T4 and TSH |
| CN108414769A (en) * | 2018-02-08 | 2018-08-17 | 江苏三联生物工程有限公司 | A kind of protein chip and preparation method thereof for heart failure marker detection |
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|---|---|---|---|---|
| WO1992021769A1 (en) * | 1991-05-30 | 1992-12-10 | Abbott Laboratories | Reagents containing a nonspecific binding blocker in ion-capture binding assays |
| CN1412320A (en) * | 2001-10-11 | 2003-04-23 | 宋克 | Joint treatment system of gene chip and its related technique |
| CN1425920A (en) * | 2002-12-26 | 2003-06-25 | 浙江大学 | Method for fixing biological macro molecule in common pattern on inorganic silicone material surface |
| CN1548549A (en) * | 2003-05-19 | 2004-11-24 | �廪��ѧ | A microparticle-based biochip system and its application |
| CN1552895A (en) * | 2003-06-04 | 2004-12-08 | 哈尔滨基太生物芯片开发有限责任公司 | Substrate surface derivatization treating technology for gene chip |
-
2007
- 2007-10-30 CN CN 200710134477 patent/CN101424682B/en active Active
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO1992021769A1 (en) * | 1991-05-30 | 1992-12-10 | Abbott Laboratories | Reagents containing a nonspecific binding blocker in ion-capture binding assays |
| CN1412320A (en) * | 2001-10-11 | 2003-04-23 | 宋克 | Joint treatment system of gene chip and its related technique |
| CN1425920A (en) * | 2002-12-26 | 2003-06-25 | 浙江大学 | Method for fixing biological macro molecule in common pattern on inorganic silicone material surface |
| CN1548549A (en) * | 2003-05-19 | 2004-11-24 | �廪��ѧ | A microparticle-based biochip system and its application |
| CN1552895A (en) * | 2003-06-04 | 2004-12-08 | 哈尔滨基太生物芯片开发有限责任公司 | Substrate surface derivatization treating technology for gene chip |
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| Title |
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| 虞伟 等.琼脂糖凝胶及片用于蛋白质分子的固定研究与表征.《第十届全军检验医学学术会议论文汇编》.2005,183-191. * |
| 鲁艳芹 等.寡核苷酸芯片制备及其杂交条件的优化.《山东大学学报（理学版）》.2004,第39卷(第3期),92-96. * |
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Denomination of invention: Biological chip base processing method Effective date of registration: 20190726 Granted publication date: 20130102 Pledgee: Agricultural Bank of China Limited by Share Ltd Wuxi science and Technology Branch Pledgor: Jiangsu Sanlian Bioengineering Co., Ltd. Registration number: 2019990000792 |
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Date of cancellation: 20200810 Granted publication date: 20130102 Pledgee: Agricultural Bank of China Limited by Share Ltd. Wuxi science and Technology Branch Pledgor: SUNLANT BIOLOGICAL ENGINEERING Co.,Ltd. Registration number: 2019990000792 |
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Address after: No. 35-305, Changjiang South Road, Xinwu District, Wuxi, Jiangsu 214111 Patentee after: Jiangsu Sanlian Bioengineering Co.,Ltd. Address before: 214101 Wuxi New District, Jiangsu Province, Xingyuan South Road, Southeast University, Wuxi Campus Patentee before: SUNLANT BIOLOGICAL ENGINEERING CO.,LTD. |
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